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Superficially bound acetylcholinesterase based on a chitosan matrix for neurotoxic compound assay by a photographic technique
Authors: Kostelník Adam | Pohanka Miroslav
Year: 2018
Type of publication: článek v odborném periodiku
Name of source: Analytical Letters
Page from-to: 1622-1632
Titles:
Language Name Abstract Keywords
cze Acetylcholinesteráza vázaná na povrchu chitosanu pro stanovení neurotoxických látek Smartphony dosáhly v posledních letech velké popularity a prosadily se i v analytické chemii, jako jednoduchý detekční systém v porovnání se standardním laboratorním vybavením. Tato práce je zaměřena na imobilizaci acetylcholinesterázy na povrch aktivované chitosanové membrány. Jako substrát byl požit alternativní substrát indoxyl acetát, z testovaných modelových inhibitorů byly použity galantamin a takrin. Měření probíhalo na 3D tištěném stojanu a fotografie byla hodnocena pomocí RGB bodů. Metoda byla dále validována za použití standardního Ellmanova spektrofotometrického testu a rovněž byla zkoumána dlouhodobá stabilita vázaného enzymu a jeho citlivost k organickým rozpouštědlům. Acetylcholinesteráza; chitosan; indoxylacetát; imobilizace
eng Superficially bound acetylcholinesterase based on a chitosan matrix for neurotoxic compound assay by a photographic technique Smartphones scored big success in the last decade and they broke into analytical chemistry as an easy detection system with comparable parameters to standard laboratory equipment. This work focuses on attachment of enzyme acetylcholinesterase (AChE) onto previously activated chitosan. Here, AChE splits alternative substrate indoxyl acetate. Conversion of substrate is blocking or slowing down by inhibitors from which galantamine and tacrine were tested here as model inhibitors with detection limits, 1.1 µM and 0.18 µM, respectively. Measurement procedure was carried out on a 3D three-dimensional printed holder and red-green-blue (RGB) channels served for digital photography evaluation. The method was validated to standard Ellman´s spectrophotometric assay. Long-term stability of immobilized enzyme as well as sensitivity to organic solvents were also tested. Acetylcholinesterase; chitosan; indoxylacetate; immobilization

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